【作者】 单保恩； 梁文杰； 任凤芝；

【Author】 ShanBao-En Liang Wen-Jie Liu Dong-Qing Ren Feng-Zhi the fourth hospital of Hebei medical university & Hebei provincial tumor hospital, Shijiazhuang, 050011, China

【机构】 河北医科大学第四医院暨河北省肿瘤医院科研中心；

【摘要】 目的:探讨北豆根有效成分诱导肿瘤细胞凋亡的作用及其作用机制。方法:北豆根干品经乙醇提取,得到北豆根醇提液,再用三步法将醇提液纯化,得到三种纯化提取物(Purified extract,PE)(PE1、2、3)。应用流式细胞技术分析了细胞周期DNA变化。经荧光染色实验和荧光显微镜观察分析了凋亡细胞形态学。用电子显微镜观察了凋亡细胞的超微结构变化。经凝胶电泳实验检测了凋亡细胞DNA 变化。结果:PE2可诱导BGC823细胞凋亡,呈明显的时效和量效关系;在低浓度PE2作用下,多数细胞被阻滞在G0／ G1期,高浓度PE2作用后,多数细胞被阻滞在S期。肿瘤细胞经PE2作用后发生细胞凋亡的形态学变化,如体积缩小,胞质浓缩,出现核固缩、核碎裂等;在电子显微镜下可见细胞胞体缩小,核染色质高度浓缩,边集于核膜,有呈半月形,有的出现核碎裂,偶见凋亡小体。细胞DNA经凝胶电泳后,出现典型的梯状电泳模式。PE2处理的癌细胞,bcl- 2基因表达降低,而bax基因表达升高。结论:PE2诱导肿瘤细胞凋亡,可通过调节bcl-2及bax基因的表达发挥其作用,这是PE2抗肿瘤作用的机制之一。该结果可促进北豆根的二次开发和临床应用。更多还原

【Abstract】 Objective: Study the effect of tumor cellular apoptosis induced by Rhizoma Menipermi Extracts-PE2 and their mechanisms. Methods: Rhizoma Menipermi extract with ethanol was purified preliminariy using three steps method. PE2 was isolated and purified farther by Pole Chromatogram. The apoptosis of tumor cells BGC823 induced by PE2 was studied by fluorescent staining, flow-cytometry (FCM), electric microscope and DNA agarose gel electrophoresis technique. Results: PE2 can induce the apoptosis of BGC823 cells with dose and time-dependent relationship. After treatment with PE2, the BGC823 cells presented some typical morphologic features and su-per-microstructural changes of apoptosis, including cell shrinkage, nclius forming outwardly acute angle promineney, chromatin concentrating on karyotheca or forming meniscus, nuclear condensation, nuclear fragmentation and formation of apoptotic bodies. Some typical subdiploid peaks before G0/G1 phase were observed. DNA agarose gel electrophoresis showed characteristic "DNA ladder" pattern. After treatment with 5 μ g/ml PE2, the cells in G0/G1 phase were blocked and increased remarkably. But the cells in S phase were blocked after treatment with 50 μ g/ml PE2. And the expression of bcl-2 was inhibited whereas the expression of bax was up-regulated. Conclusion: Rhizoma Menispermi Extract PE2 induces cellular apoptosis, which are important mechanisms of anti-tumor effect. Futher purification and study for their mechanisms can produce the scientific evidence for developing and manufacturing new antitumor drugs.更多还原